De Maria R, Cifone M G, Trotta R, Rippo M R, Festuccia C, Santoni A, Testi R
Department of Experimental Medicine and Biochemical Sciences, University of Rome Tor Vergata, Italy.
J Exp Med. 1994 Nov 1;180(5):1999-2004. doi: 10.1084/jem.180.5.1999.
The expression and function of CD69, a member of the natural killer cell gene complex family of signal transducing receptors, was investigated on human monocytes. CD69 was found expressed on all peripheral blood monocytes, as a 28- and 32-kD disulfide-linked dimer. Molecular cross-linking of CD69 receptors induced extracellular Ca2+ influx, as revealed by flow cytometry. CD69 cross-linking resulted also in phospholipase A2 activation, as detected by in vivo arachidonic acid release measurement from intact cells and by direct in vitro measurement of enzymatic activity using radiolabeled phosphatidylcholine vesicles. Prostaglandin E 2 alpha, 6-keto-prostaglandin F 1 alpha, and leukotriene B4 were detected by radioimmunoassay in supernatants from CD69-stimulated monocytes, suggesting the activation of both cyclooxygenase and lipoxygenase pathways after CD69 stimulation. CD69 cross-linking, moreover, was able to induce strong nitric oxide (NO) production from monocytes, as detected by accumulation of NO oxydixed derivatives, and cyclic GMP. It is important to note that NO generation was responsible for CD69-mediated increase in spontaneous cytotoxicity against L929 murine transformed fibroblast cell line and induction of redirected cytotoxicity towards P815 FcRII+ murine mastocytoma cell line. These data indicate that CD69 can act as a potent stimulatory molecule on the surface of human peripheral blood monocytes.
对人单核细胞研究了信号转导受体自然杀伤细胞基因复合体家族成员CD69的表达及功能。发现CD69在所有外周血单核细胞上均有表达,为一种28-kD和32-kD的二硫键连接二聚体。流式细胞术显示,CD69受体的分子交联诱导细胞外Ca2+内流。CD69交联还导致磷脂酶A2激活,这通过完整细胞中花生四烯酸释放的体内测量以及使用放射性标记磷脂酰胆碱囊泡对酶活性的直接体外测量得以检测。通过放射免疫分析法在CD69刺激的单核细胞上清液中检测到前列腺素E2α、6-酮-前列腺素F1α和白三烯B4,提示CD69刺激后环氧化酶和脂氧化酶途径均被激活。此外,CD69交联能够诱导单核细胞产生大量一氧化氮(NO),这通过NO氧化衍生物和环磷酸鸟苷的积累得以检测。值得注意的是,NO的产生是CD69介导的对L929小鼠转化成纤维细胞系自发细胞毒性增加以及对P815 FcRII+小鼠肥大细胞瘤细胞系重定向细胞毒性诱导的原因。这些数据表明,CD69可作为人外周血单核细胞表面的一种强效刺激分子。